CN112136962B - Enzymolysis wheat protein powder and preparation method thereof - Google Patents
Enzymolysis wheat protein powder and preparation method thereof Download PDFInfo
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- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
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Abstract
The invention belongs to the technical field of enzymolysis, and particularly relates to enzymolysis wheat protein powder and a preparation method thereof. The preparation method of the enzymatic hydrolysis wheat protein powder provided by the invention comprises the steps of preparing wheat gluten into slurry, adding a stabilizer, and homogenizing to obtain a homogeneous liquid; adjusting the pH of the homogenized solution, heating, adding complex enzyme, performing enzymolysis, performing enzyme inactivation, cooling to 25 deg.C to obtain enzymolysis wheat protein solution, and spray drying the obtained enzymolysis wheat protein solution. The preparation method of the enzymolysis wheat protein powder provided by the invention can obviously improve the hydrolysis degree, solubility and digestibility of the wheat protein.
Description
Technical Field
The invention belongs to the technical field of enzymolysis, and particularly relates to enzymolysis wheat protein powder and a preparation method thereof.
Background
The wheat protein powder is a byproduct generated in the production process of wheat starch, is a vegetable protein source, is rich in nutrition, contains 15 kinds of amino acids necessary for a human body, contains more hydrophobic and uncharged amino acid contents, is difficult to adapt to processing requirements, and is limited in application to a certain extent. The main component of the wheat protein powder is protein, the content of other amino acids in the wheat protein powder protein is relatively high except that the content of lysine is relatively low, particularly the content of glutamic acid accounts for 30 percent of the content of the whole amino acid, and the wheat protein powder protein is very beneficial to the intestinal health of animals. Therefore, the natural plant protein source is regarded as important in feed production, and is particularly applied to pet and high-grade animal feed.
The wheat protein powder is prepared by physical method, enzyme method, chemical method and gene engineering method. Among them, the chemical method may cause harmful effects in nutrition and toxicology while obtaining expected effects, and the genetic engineering method is difficult to be applied industrially due to long technical cycle. The enzyme method is widely regarded as having advantages of strong controllability, moderate conditions, less energy consumption, strong specificity, less byproducts and the like. However, the existing enzymolysis process of wheat protein has the problems of low hydrolysis degree, unsatisfactory solubility of the prepared enzymolysis wheat protein powder, low digestibility and the like.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide enzymolysis wheat protein powder and a preparation method thereof. The preparation method of the enzymolysis wheat protein powder provided by the invention can obviously improve the hydrolysis degree, solubility and digestibility of the wheat protein and broaden the application range of the enzymolysis wheat protein powder.
The technical scheme of the invention is as follows:
a preparation method of enzymolysis wheat protein powder comprises the following steps:
s1 adding water into wheat gluten to obtain slurry with mass concentration of 10-15%, adding stabilizer, homogenizing under 10-20Mpa for 2 times to obtain homogenized solution;
s2, adjusting the pH value of the homogenized liquid obtained in the step S1 to 7-9, heating to 40-45 ℃, adding a complex enzyme under stirring at the stirring speed of 200-300r/min, wherein the adding amount of the complex enzyme is 0.3-0.5 per mill of the mass of the homogenized liquid obtained in the step S1, carrying out enzymolysis for 1-3h, heating to 80-95 ℃ for carrying out enzyme inactivation for 8-20min, and cooling to 25 ℃ to obtain an enzymolysis wheat protein liquid;
s3, spray drying the enzymolysis wheat protein liquid obtained in the step S2 to obtain the enzyme-hydrolyzed wheat protein liquid.
Further, the stabilizer in the step S1 is composed of sodium alginate and chitosan oligosaccharide in a mass ratio of 5-8: 2-3.
Further, the stabilizer in the step S1 is composed of sodium alginate and chitosan oligosaccharide in a mass ratio of 7: 2.
Further, the molecular weight of the chitosan oligosaccharide is 2000-3000.
Further, the complex enzyme in the step S2 is composed of modified chymotrypsin, cellulase and amylase according to the mass ratio of 12-14:5-8: 1-2.
Further, the compound enzyme is composed of modified chymotrypsin, cellulase and amylase according to the mass ratio of 13:6: 2.
Further, the modified chymotrypsin is prepared from the following components in parts by mass:
3-5 parts of chymotrypsin, 2-4 parts of polyvinylpyrrolidone, 10-15 parts of maltodextrin, 1-3 parts of polyvinyl alcohol and 150 parts of water.
Further, the modified chymotrypsin is prepared from the following components in parts by mass:
4 parts of chymotrypsin, 3 parts of polyvinylpyrrolidone, 12 parts of maltodextrin, 2 parts of polyvinyl alcohol and 128 parts of water.
The invention also aims to provide the enzymolysis wheat protein powder prepared by the preparation method of the enzymolysis wheat protein powder.
The preparation method of the enzymatic hydrolysis wheat protein powder provided by the invention comprises the steps of preparing wheat gluten into slurry, adding a stabilizer, homogenizing, adding a complex enzyme, carrying out enzymatic hydrolysis and the like. The preparation method of the enzymolysis wheat protein powder provided by the invention can obviously improve the hydrolysis degree, solubility and digestibility of the wheat protein and broaden the application range of the enzymolysis wheat protein powder.
According to the invention, the stabilizing agent is added into the slurry for homogenization, so that the uniformity of the slurry can be improved, the stretching of a protein molecular structure is facilitated, and the solubility of wheat protein is enhanced. According to the invention, the complex enzyme compounded by the modified chymotrypsin, the cellulase and the amylase according to a certain mass ratio can generate an excellent enzymolysis effect, further improve the solubility of the wheat protein, release various types of physiologically active peptides from the wheat protein, improve the protein digestion rate and be beneficial to the absorption and utilization of the wheat protein. In the invention, the modified chymotrypsin prepared from chymotrypsin, polyvinylpyrrolidone, maltodextrin and polyvinyl alcohol can be isolated from cellulase and amylase, so that the complex enzyme has more excellent enzymolysis effect.
Compared with the prior art, the invention has the following advantages:
(1) the preparation method of the enzymolysis wheat protein powder provided by the invention can obviously improve the hydrolysis degree and solubility of the wheat protein and expand the application range of the enzymolysis wheat protein powder.
(2) The preparation method of the enzymolysis wheat protein powder provided by the invention is simple and easy to operate, and is easy for industrial application.
(3) The enzymolysis wheat protein powder prepared by the preparation method of the enzymolysis wheat protein powder provided by the invention has high digestibility and is beneficial to absorption and utilization.
Detailed Description
The present invention is further described in the following description of the specific embodiments, which is not intended to limit the invention, but various modifications and improvements can be made by those skilled in the art according to the basic idea of the invention, within the scope of the invention, as long as they do not depart from the basic idea of the invention.
In the present invention, the starting materials are commercially available unless otherwise specified. For example, wheat gluten can be purchased from Jiangsu Chengji Biotechnology Co., Ltd, article number: 1031; sodium alginate is available from Guangdong Hua Zhan pharmaceutical Co., Ltd, cat #: 1002; chymotrypsin is available from biotechnology limited company of wakamaike, beijing, cat #: 831885, respectively; cellulases are available from shanxi to dershengtong food ltd, cat #: 6875; amylases are available from shanxi to dershengton food ltd, cat #: 235; polyvinylpyrrolidone is available from Guangdong Yuebeautified industries, Inc., cat #: k 30; maltodextrin is available from wu yi chemical technology ltd, suzhou under the cat code: 058; polyvinyl alcohol is available from inner mongolia kadu new materials science and technology ltd, cat #: 2488.
example 1 preparation method of enzymolysis wheat protein powder
The preparation method of the enzymolysis wheat protein powder comprises the following steps:
s1, taking wheat gluten, adding water to prepare slurry with the mass concentration of 10%, adding a stabilizer, wherein the stabilizer consists of sodium alginate and chitosan oligosaccharide with the molecular weight of 2000 according to the mass ratio of 5:3, homogenizing for 2 times under 10Mpa to obtain homogenized liquid;
s2, adjusting the pH value of the homogenized liquid obtained in the step S1 to 7, heating to 40 ℃, adding a complex enzyme under stirring at a stirring speed of 200r/min, wherein the complex enzyme consists of modified chymotrypsin, cellulase and amylase according to a mass ratio of 12:8:2, the adding amount of the complex enzyme is 0.3 per mill of the mass of the homogenized liquid obtained in the step S1, the enzymolysis time is 1h, heating to 80 ℃ for enzyme inactivation, the enzyme inactivation time is 8min, and cooling to 25 ℃ to obtain an enzymolysis wheat protein liquid;
s3, spray drying the enzymolysis wheat protein liquid obtained in the step S2 to obtain the enzyme-hydrolyzed wheat protein liquid.
The modified chymotrypsin is prepared from the following components in parts by mass:
3 parts of chymotrypsin, 2 parts of polyvinylpyrrolidone, 10 parts of maltodextrin, 1 part of polyvinyl alcohol and 120 parts of water.
The preparation method of the modified chymotrypsin comprises the following steps:
(1) mixing polyvinylpyrrolidone, maltodextrin and polyvinyl alcohol, adding water, and stirring to completely dissolve to obtain a mixed solution;
(2) adding chymotrypsin into the mixed solution obtained in the step (1), and stirring until the chymotrypsin is completely dissolved to obtain an enzyme mixed solution;
(3) and (3) spray drying the enzyme mixed solution obtained in the step (2) to obtain the enzyme-containing water-soluble chitosan.
Example 2 preparation method of enzymolysis wheat protein powder
The preparation method of the enzymolysis wheat protein powder comprises the following steps:
s1, taking wheat gluten, adding water to prepare slurry with the mass concentration of 15%, adding a stabilizer, wherein the stabilizer consists of sodium alginate and chitosan oligosaccharide with the molecular weight of 3000 according to the mass ratio of 8:2, homogenizing for 2 times under 20Mpa to obtain a homogeneous solution;
s2, adjusting the pH value of the homogenized liquid obtained in the step S1 to 9, heating to 45 ℃, adding a complex enzyme under stirring at a stirring speed of 300r/min, wherein the complex enzyme consists of modified chymotrypsin, cellulase and amylase according to a mass ratio of 14:5:1, the adding amount of the complex enzyme is 0.5 per mill of the mass of the homogenized liquid obtained in the step S1, the enzymolysis time is 3 hours, heating to 95 ℃ for enzyme inactivation, the enzyme inactivation time is 20min, and cooling to 25 ℃ to obtain an enzymolysis wheat protein liquid;
s3, spray drying the enzymolysis wheat protein liquid obtained in the step S2 to obtain the enzyme-hydrolyzed wheat protein liquid.
The modified chymotrypsin is prepared from the following components in parts by mass:
5 parts of chymotrypsin, 4 parts of polyvinylpyrrolidone, 15 parts of maltodextrin, 3 parts of polyvinyl alcohol and 150 parts of water.
The preparation method of the modified chymotrypsin is similar to that of example 1.
Example 3 preparation method of enzymolysis wheat protein powder
The preparation method of the enzymolysis wheat protein powder comprises the following steps:
s1, taking wheat gluten, adding water to prepare slurry with the mass concentration of 14%, adding a stabilizer, wherein the stabilizer consists of sodium alginate and chitosan oligosaccharide with the molecular weight of 2500 according to the mass ratio of 7:2, homogenizing for 2 times under 12Mpa to obtain a homogeneous solution;
s2, adjusting the pH value of the homogenized liquid obtained in the step S1 to 8, heating to 42 ℃, adding a complex enzyme under stirring at a stirring speed of 240r/min, wherein the complex enzyme consists of modified chymotrypsin, cellulase and amylase according to a mass ratio of 13:6:2, the adding amount of the complex enzyme is 0.4 per mill of the mass of the homogenized liquid obtained in the step S1, the enzymolysis time is 1.5h, heating to 90 ℃ for enzyme inactivation, the enzyme inactivation time is 12min, and cooling to 25 ℃ to obtain an enzymolysis wheat protein liquid;
s3, spray drying the enzymolysis wheat protein liquid obtained in the step S2 to obtain the enzyme-hydrolyzed wheat protein liquid.
The modified chymotrypsin is prepared from the following components in parts by mass:
4 parts of chymotrypsin, 3 parts of polyvinylpyrrolidone, 12 parts of maltodextrin, 2 parts of polyvinyl alcohol and 128 parts of water.
The preparation method of the modified chymotrypsin is similar to that of example 1.
Comparative example 1 preparation method of enzymolysis wheat protein powder
The preparation method of the enzymolysis wheat protein powder comprises the following steps:
s1, taking wheat gluten, adding water to prepare slurry with the mass concentration of 14%;
s2, adjusting the pH value of the slurry obtained in the step S1 to 8, heating to 42 ℃, adding a complex enzyme under stirring at a stirring speed of 240r/min, wherein the complex enzyme consists of modified chymotrypsin, cellulase and amylase according to a mass ratio of 13:6:2, the adding amount of the complex enzyme is 0.4 per mill of the mass of the homogenized liquid obtained in the step S1, the enzymolysis time is 1.5h, heating to 90 ℃ for enzyme inactivation, the enzyme inactivation time is 12min, and cooling to 25 ℃ to obtain an enzymolysis wheat protein liquid;
s3, spray drying the enzymolysis wheat protein liquid obtained in the step S2 to obtain the enzyme-hydrolyzed wheat protein liquid.
The modified chymotrypsin is prepared from the following components in parts by mass:
4 parts of chymotrypsin, 3 parts of polyvinylpyrrolidone, 12 parts of maltodextrin, 2 parts of polyvinyl alcohol and 128 parts of water.
The preparation method of the modified chymotrypsin is similar to that of example 1.
Comparative example 2 preparation method of enzymolysis wheat protein powder
The preparation method of the enzymolysis wheat protein powder comprises the following steps:
s1, taking wheat gluten, adding water to prepare slurry with the mass concentration of 14%, adding a stabilizer, wherein the stabilizer consists of sodium alginate and chitosan oligosaccharide with the molecular weight of 2500 according to the mass ratio of 7:2, homogenizing for 2 times under 12Mpa to obtain a homogeneous solution;
s2, adjusting the pH value of the homogenized liquid obtained in the step S1 to 8, heating to 42 ℃, adding a complex enzyme under stirring at a stirring speed of 240r/min, wherein the complex enzyme consists of modified chymotrypsin, cellulase and amylase according to a mass ratio of 1:1:1, the adding amount of the complex enzyme is 0.4 per mill of the mass of the homogenized liquid obtained in the step S1, the enzymolysis time is 1.5h, heating to 90 ℃ for enzyme inactivation, the enzyme inactivation time is 12min, and cooling to 25 ℃ to obtain an enzymolysis wheat protein liquid;
s3, spray drying the enzymolysis wheat protein liquid obtained in the step S2 to obtain the enzyme-hydrolyzed wheat protein liquid.
The modified chymotrypsin is prepared from the following components in parts by mass:
4 parts of chymotrypsin, 3 parts of polyvinylpyrrolidone, 12 parts of maltodextrin, 2 parts of polyvinyl alcohol and 128 parts of water.
The preparation method of the modified chymotrypsin is similar to that of example 1.
Comparative example 3 preparation method of enzymolysis wheat protein powder
The preparation method of the enzymolysis wheat protein powder comprises the following steps:
s1, taking wheat gluten, adding water to prepare slurry with the mass concentration of 14%, adding a stabilizer, wherein the stabilizer consists of sodium alginate and chitosan oligosaccharide with the molecular weight of 2500 according to the mass ratio of 7:2, homogenizing for 2 times under 12Mpa to obtain a homogeneous solution;
s2, adjusting the pH value of the homogenized liquid obtained in the step S1 to 8, heating to 42 ℃, adding a complex enzyme under stirring at a stirring speed of 240r/min, wherein the complex enzyme consists of chymotrypsin, cellulase and amylase according to a mass ratio of 13:6:2, the adding amount of the complex enzyme is 0.4 per mill of the mass of the homogenized liquid obtained in the step S1, the enzymolysis time is 1.5h, heating to 90 ℃ for enzyme inactivation, the enzyme inactivation time is 12min, and cooling to 25 ℃ to obtain an enzymolysis wheat protein liquid;
s3, spray drying the enzymolysis wheat protein liquid obtained in the step S2 to obtain the enzyme-hydrolyzed wheat protein liquid.
Test example I measurement of hydrolysis degree
1. Test materials: the enzymolytic wheat protein solutions prepared in examples 1, 2 and 3.
2. The test method comprises the following steps:
placing 5.00mL of enzymolysis wheat protein liquid in a small beaker, adding 20mL of distilled water without carbon dioxide, carrying out magnetic stirring, dropwise adding 0.1mol/L NaOH standard solution until the pH value is 8.2, then adding 10mL of neutralized formaldehyde solution, and recording the milliliter number of 0.1mol/L NaOH solution consumed when the pH value is titrated to 9.2; blank tests were performed. The amino nitrogen of the protein in the enzymolysis wheat protein liquid is as follows:
enzymolysis wheat protein liquid amino nitrogen (g) [ (A-B) × NxV ═ N1×k]/V
Wherein A is the volume of the sodium hydroxide standard solution consumed by the titration sample, mL; b is the volume of the blank sodium hydroxide standard solution consumed by titration, mL; n is the concentration of the sodium hydroxide standard solution, mol/L; k is 1mol/L sodium hydroxide 1mL corresponding to the mass of nitrogen, g, k is 0.014; v is the volume of the sample, mL; v1Is the total volume of the hydrolysate, mL.
Degree of hydrolysis ═ amino nitrogen/total nitrogen x 100%.
3. And (3) test results:
the test results are shown in table 1.
Table 1: measurement result of degree of hydrolysis
| Group of | Example 1 | Example 2 | Example 3 |
| Degree of hydrolysis (%) | 18.62 | 25.43 | 32.15 |
As can be seen from Table 1, the enzymatic hydrolyzed wheat proteins prepared in examples 1-3 of the present invention have a high degree of hydrolysis. Among them, the enzymolysis wheat protein of the embodiment 3 has the highest hydrolysis degree, and is the best embodiment of the invention.
Test example II measurement of solubility
1. Test materials: the enzymolysis wheat protein powder prepared in the example 3, the comparative example 1, the comparative example 2 and the comparative example 3.
2. The test method comprises the following steps:
the solubility of the prepared enzymolysis wheat protein powder is determined by adopting a forskolin reagent method.
3. And (3) test results:
the test results are shown in table 2.
Table 2: results of solubility measurement
| Group of | Example 3 | Comparative example 1 | Comparative example 2 | Comparative example 3 |
| Solubility (%) | 86.23 | 74.16 | 76.85 | 69.37 |
As can be seen from Table 2, the solubility of the enzymatic hydrolyzed wheat protein powder prepared in example 3 of the present invention is significantly higher than that of the enzymatic hydrolyzed wheat protein powders prepared in comparative examples 1-3. The method for preparing the enzymolysis wheat protein powder has the advantage that the enzymolysis wheat protein powder obtained by the method for preparing the enzymolysis wheat protein powder has good dissolvability.
Test example three measurement of in vitro digestibility
1. Test materials: the enzymolysis wheat protein powder prepared in the example 3, the comparative example 2 and the comparative example 3.
2. The test method comprises the following steps:
weighing 1.0g of test material, dissolving in 80mL of HCl solution with concentration of 0.05mol/L (pH of 1.5), setting a blank control group, placing on a shaker at 39 ℃, preheating for 5min, adding a certain amount of pepsin (enzyme: substrate: 1:100, mass ratio), digesting for 3h, adjusting pH of the zymolyte solution to 6.8 by NaOH solution with concentration of 1mol/L, adding a certain amount of trypsin (enzyme: substrate: 1:20, mass ratio), digesting for 2h, taking out the solution, centrifuging by a refrigerated centrifuge (10000r/min, 4 ℃, 10min), and measuring protein content of the centrifuged supernatant by a half Kjeldahl method. The ratio of the difference value of the protein content of the experimental group supernatant and the protein content of the blank control group supernatant to the total protein content in the enzymolysis sample is the digestibility of the protein.
3. And (3) test results:
the test results are shown in table 3.
Table 3: results of in vitro digestibility determination
| Group of | Example 3 | Comparative example 2 | Comparative example 3 |
| In vitro digestibility (%) | 88.16 | 79.31 | 68.27 |
As can be seen from table 3, the in vitro digestibility of the enzymatic hydrolyzed wheat protein powder prepared in example 3 of the present invention is significantly higher than the in vitro digestibility of the enzymatic hydrolyzed wheat protein powders prepared in comparative examples 2 and 3. The method for preparing the enzymatic hydrolysis wheat protein powder has the advantages that the digestion rate of the enzymatic hydrolysis wheat protein powder is high, and the enzymatic hydrolysis wheat protein powder is easy to absorb and utilize.
Claims (7)
1. A preparation method of enzymolysis wheat protein powder is characterized by comprising the following steps:
s1 adding water into wheat gluten to obtain slurry with mass concentration of 10-15%, adding stabilizer, homogenizing under 10-20Mpa for 2 times to obtain homogenized solution;
s2, adjusting the pH value of the homogenized liquid obtained in the step S1 to 7-9, heating to 40-45 ℃, adding a complex enzyme under stirring at the stirring speed of 200-300r/min, wherein the adding amount of the complex enzyme is 0.3-0.5 per mill of the mass of the homogenized liquid obtained in the step S1, carrying out enzymolysis for 1-3h, heating to 80-95 ℃ for carrying out enzyme inactivation for 8-20min, and cooling to 25 ℃ to obtain an enzymolysis wheat protein liquid;
s3, spray drying the enzymolysis wheat protein liquid obtained in the step S2 to obtain the enzymolysis wheat protein liquid;
the compound enzyme in the step S2 consists of modified chymotrypsin, cellulase and amylase according to the mass ratio of 12-14:5-8: 1-2;
the modified chymotrypsin is prepared from the following components in parts by mass:
3-5 parts of chymotrypsin, 2-4 parts of polyvinylpyrrolidone, 10-15 parts of maltodextrin, 1-3 parts of polyvinyl alcohol and 150 parts of water.
2. The preparation method of the enzymatic hydrolysis wheat protein powder as claimed in claim 1, wherein the stabilizer in the step S1 is composed of sodium alginate and chitosan oligosaccharide in a mass ratio of 5-8: 2-3.
3. The preparation method of the enzymatic hydrolysis wheat protein powder as claimed in claim 2, wherein the stabilizer in the step S1 is composed of sodium alginate and chitosan oligosaccharide in a mass ratio of 7: 2.
4. The method for preparing enzymatic hydrolyzed wheat protein powder as claimed in claim 2, wherein the molecular weight of the chitosan oligosaccharide is 2000-3000.
5. The preparation method of the enzymatic hydrolysis wheat protein powder as claimed in claim 1, wherein the complex enzyme is composed of modified chymotrypsin, cellulase and amylase according to the mass ratio of 13:6: 2.
6. The preparation method of the enzymatic hydrolysis wheat protein powder as claimed in claim 1, wherein the modified chymotrypsin is prepared from the following components in parts by weight:
4 parts of chymotrypsin, 3 parts of polyvinylpyrrolidone, 12 parts of maltodextrin, 2 parts of polyvinyl alcohol and 128 parts of water.
7. The enzymatic hydrolyzed wheat protein powder prepared by the preparation method of the enzymatic hydrolyzed wheat protein powder according to any one of claims 1 to 6.
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| CN104605137A (en) * | 2015-02-15 | 2015-05-13 | 江苏康科食品工程技术有限公司 | Preparation method of wheat protolysate |
| CN106417891A (en) * | 2015-08-05 | 2017-02-22 | 江苏爱福特科技开发有限公司 | Method for preparing gluten powder hydrolysate by utilizing enzymatic method |
| CN111670995A (en) * | 2020-06-08 | 2020-09-18 | 颜如玉医药科技有限公司 | Preparation method of wheat protein peptide |
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| CN104605137A (en) * | 2015-02-15 | 2015-05-13 | 江苏康科食品工程技术有限公司 | Preparation method of wheat protolysate |
| CN106417891A (en) * | 2015-08-05 | 2017-02-22 | 江苏爱福特科技开发有限公司 | Method for preparing gluten powder hydrolysate by utilizing enzymatic method |
| CN111670995A (en) * | 2020-06-08 | 2020-09-18 | 颜如玉医药科技有限公司 | Preparation method of wheat protein peptide |
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| 小麦面筋蛋白研究进展;王岸娜等;《粮油食品科技》;20171231;第25卷(第5期);19-23 * |
| 湿热-胶体改性对小麦面筋蛋白功能性质的影响;许慧等;《食品工业科技》;20111231;第32卷(第7期);147-149、153 * |
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